Abstract The cranial neural crest (CNC) cells, the migrating multipotent cells giving rise to different derivatives composing the majority of the craniofacial structures including bone, cartilage, neurons, glia, smooth muscle, melanocytes and adipocytes. Abnormal CNC development results in the most common congenital human birth defects like cleft lip/palate, craniosynostosis, and genetic syndromes, giving a large impact of the life of patients and their families. Therefore, the objective of our proposed study is to understand the molecular regulatory mechanism regulating CNC development. Key Hippo signaling roles have been broadly observed in development and diseases involving many different tissues and organs. However, Hippo's role in craniofacial development is still poorly understood. Wnt signaling is known to play a pivotal role in CNC development and cross talks with Hippo signaling in some context like in heart, whereas there is a knowledge gap of Hippo and Wnt interaction in craniofacial development. We have obtained promising preliminary data indicate that Hippo signaling interacts with Wnt signaling and plays a critical role in regulating CNC-derived craniofacial development. In this proposed study, we will dissect the important roles of Hippo-Yap pathway in CNC cells migration and differentiation, and how Hippo and Wnt pathways cross talk to regulate CNC derived craniofacial development. The goal of the proposed studies is to elucidate the molecular regulatory mechanisms underlying CNC development and help to develop novel therapeutic strategies and diagnostic tools for craniofacial defects.